Bioenergetic consequences of protein overexpression in Saccharomyces cerevisiae

C, Gopal; Broad, David; Lloyd, David

doi:10.1007/bf00264005

Cited by 27 publications

(25 citation statements)

References 19 publications

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“…For recombinant S. cerevisiae strains, reductions in the maximum specific growth rate, biomass yield, respiratory capacity, and stability of the recombinant plasmid, due to heterologous gene expression, have been observed (Da Silva and Bailey, 1991;Dequin and Barre, 1994;Gopal et al, 1989;Giuseppin et al, 1993;Janes et al, 1990;Marquet et al, 1987;Nacken et al, 1996;Sardonini and DiBiasio, 1987;Srienc et al, 1986). A decrease in the flux through glycolysis of the host strain (Snoep et al, 1995;Van Hoek et al, 1998) and an increase in the maintenance energy requirement (Bhattacharya and Dubey, 1995;Stouthamer and Van Verseveld, 1987) have also been observed.…”

Section: Introductionmentioning

confidence: 87%

“…The energetic cost for the synthesis of recombinant protein to the equivalent of 0.2% cellular protein during growth on glucose could not be translated directly into the 11% reduction in the maximum specific growth rate of the Y294 [PGK1-XYN] strain (Table II). Reduced production levels for heterologous gene expression, compared with homologous gene expression, have also been observed for production from the same expression system (Chen et al, 1984;Gopal et al, 1989), apparently due to a higher energetic demand for heterologous gene expression (Gopal et al, 1989).…”

Section: Heterologous Gene Expressionmentioning

confidence: 94%

“…The metabolic burden of recombinant gene expression has been associated with the allocation of cellular resources to plasmid-related activities (Gu et al, 1996;Gopal et al, 1989;Peretti and Bailey, 1987;Ryan and Parulekar, 1991), either for plasmid replication or the production of the cloned gene product (Bailey, 1993;Bentley et al, 1990;Da Silva and Bailey, 1991;Ryan and Parulekar, 1991). The effect of cloned gene expression has been associated with either the energetic cost of extra protein synthesis (Bailey, 1993;Gopal et al, 1989;Snoep et al, 1995) or the competitive effect of extra protein synthesis (Gopal et al, 1989;Shuster, 1989;Snoep et al, 1995;Van der Aar et al, 1992).…”

Section: Introductionmentioning

confidence: 98%

“…The effect of cloned gene expression has been associated with either the energetic cost of extra protein synthesis (Bailey, 1993;Gopal et al, 1989;Snoep et al, 1995) or the competitive effect of extra protein synthesis (Gopal et al, 1989;Shuster, 1989;Snoep et al, 1995;Van der Aar et al, 1992). The "dilution" of native proteins (i.e., the reduction of the activity of native proteins by recombinant gene expression) was also proposed as a major mechanism causing a decrease in the flux through glycolysis and a decrease in the maximum specific growth rate of the host organism (Snoep et al, 1995;Van Hoek et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

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The metabolic burden of the PGK1 and ADH2 promoter systems for heterologous xylanase production by Saccharomyces cerevisiae in defined medium

Görgens

Zyl

Knoetze

et al. 2001

Biotech & Bioengineering

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Five recombinant S. cerevisiae strains were cultivated under identical conditions to quantify the molecular basis of the metabolic burden of heterologous gene expression, and to evaluate mechanisms for the metabolic burden. Two recombinant S. cerevisiae strains, producing Trichoderma reesei xylanase II under control of either the PGK1 or ADH2 promoters, were compared quantitatively with three references strains, where either the heterologous xylanase II (XYN2) gene, or the heterologous gene and the promoter and terminator were omitted from the recombinant plasmid. Neither the replication of multiple copies of the 2-microm-based YEp352 plasmid nor the replication the foreign XYN2 gene represented a metabolic burden to the cell, as the growth of the host organism was not affected. The inclusion of a glycolytic promoter on the recombinant plasmid, however, reduced the maximum specific growth rate (12% to 15%), biomass yield on glucose (8% to 11%), and specific glucose consumption rate (6% to 10%) of the recombinant strains. The presence of the heterologous XYN2 gene on the recombinant plasmid caused a further reduction in the maximum specific growth rate (11% to 14%), biomass yield (4%), and specific glucose consumption rate (12%) of the host strain during active gene expression, which was dictated by the regulatory characteristics of the promoter utilized. The metabolic effect of foreign gene expression was disproportionally large, with respect to on the amount of heterologous protein produced. This was most likely due to an increased energetic demand for the expression of a foreign gene and/or a competition for limiting amounts of transcription or translation factors, biosynthetic precursors or metabolic energy.

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Section: Introductionmentioning

confidence: 87%

Section: Heterologous Gene Expressionmentioning

confidence: 94%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The metabolic burden of the PGK1 and ADH2 promoter systems for heterologous xylanase production by Saccharomyces cerevisiae in defined medium

Görgens

Zyl

Knoetze

et al. 2001

Biotech & Bioengineering

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“…As shown in Fig. 2 and Table 2, the host strain gained the highest cell mass of 7AE3 g l )1 probably due to the low metabolic burden to express foreign proteins (Gopal et al 1989;Janes et al 1990). In ethanol production, however, three recombinant strains (S. cerevisiae L2612, L2612:YIpXK and L2612:dISXK) produced twofold higher ethanol concentrations (9AE6-11AE6 g l )1 ), twofold higher ethanol yields (0AE22-0AE27) and 5AE3-to 6AE0-fold higher overall ethanol productivities (0AE16-0AE18 g l )1 h )1 ) as compared with the host strain values.…”

Section: Discussionmentioning

confidence: 97%

Effects of xylulokinase activity on ethanol production from d-xylulose by recombinant Saccharomyces cerevisiae

et al. 2003

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Aims: Recombinant Saccharomyces cerevisiae strains harbouring different levels of xylulokinase (XK) activity and effects of XK activity on utilization of xylulose were studied in batch and fed‐batch cultures. Methods and Results: The cloned xylulokinase gene (XKS1) from S. cerevisiae was expressed under the control of the glyceraldehyde 3‐phosphate dehydrogenase promoter and terminator. Specific xylulose consumption rate was enhanced by the increased specific XK activity, resulting from the introduction of the XKS1 into S. cerevisiae. In batch and fed‐batch cultivations, the recombinant strains resulted in twofold higher ethanol concentration and 5·3‐ to six‐fold improvement in the ethanol production rate compared with the host strain S. cerevisiae. Conclusions: An effective conversion of xylulose to xylulose 5‐phosphate catalysed by XK in S. cerevisiae was considered to be essential for the development of an efficient and accelerated ethanol fermentation process from xylulose. Significance and Impact of the Study: Overexpression of the XKS1 gene made xylulose fermentation process accelerated to produce ethanol through the pentose phosphate pathway.

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Macroscopic modeling of the growth and substrate consumption of wild type and genetically modified Pichia pastoris

Joseph,

Akkermans,

Van Impe

2023

Biotechnology Journal

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Pichia pastoris is a popular yeast platform to generate several industrially relevant products which have applications in a wide range of sectors. The complexities in the processes due to the addition of a foreign gene are not widely explored. Since these complexities can be dependent on the strain characteristics, promoter, and type of protein produced, it is vital to investigate the growth and substrate consumption patterns of the host to facilitate customized process optimization. In this study, the growth rates of P. pastoris GS115 wild type (WT) and genetically modified (GM) strains grown on glycerol and methanol in batch cultivation mode were estimated and the model providing the best representation of the true growth kinetics based on substrate consumption was identified. It was observed that the growth of P. pastoris exhibits Haldane kinetics on glycerol rather than the most commonly used Monod kinetics due to the inability of the latter to describe growth inhibition at high concentrations of glycerol. Whereas, the cardinal parameter model, a newly proposed model for this application, was found to be the best fitting to describe the growth of P. pastoris on methanol due to its ability to describe methanol toxicity. Interestingly, the findings from this study concluded that in both substrates, the genetically engineered strain exhibited a higher growth rate compared to the WT strain. Such an observation has not been established yet in other published works, indicating an opportunity to further optimize the carbon source feeding strategies when the host is grown in fed‐batch mode.

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Bioenergetic consequences of protein overexpression in Saccharomyces cerevisiae

Cited by 27 publications

References 19 publications

The metabolic burden of the PGK1 and ADH2 promoter systems for heterologous xylanase production by Saccharomyces cerevisiae in defined medium

The metabolic burden of the PGK1 and ADH2 promoter systems for heterologous xylanase production by Saccharomyces cerevisiae in defined medium

Effects of xylulokinase activity on ethanol production from d-xylulose by recombinant Saccharomyces cerevisiae

Macroscopic modeling of the growth and substrate consumption of wild type and genetically modified Pichia pastoris

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